基于UPLC-Triple-TOF MS/MS对巴氏杀菌乳中磷脂成分的分析  

Identifying and Quantifying Phospholipids in Pasteurized Milk Using UPLC-Triple-TOF MS/MS

作　　者：曹雪 任皓威[1] 王筱迪 许铵颀 商佳琦 刘宁[1] CAO Xue;REN Haowei;WANG Xiaodi;XU Anqi;SHANG Jiaqi;LIU Ning(Key Laboratory of Dairy Science,Ministry of Education,College of Food Science,Northeast Agricultural University,Harbin 150030,China)

机构地区：[1]乳品科学教育部重点实验室,东北农业大学食品学院,黑龙江哈尔滨150030

出　　处：《食品科学》2021年第4期181-189,共9页Food Science

基　　金：“十三五”国家重点研发计划重点专项(2018YFC1604304-01)。

摘　　要：基于超高效液相色谱-串联四极杆飞行时间高分辨质谱(ultra-high performance liquid chromatographytriple quadrupole-time of flight tandem mass spectrometry,UPLC-Triple-TOF MS/MS)法对巴氏杀菌乳中的磷脂成分进行分析鉴定。采用色谱柱ACQUITY UPLC^®BEH C18(2.1 mm×100 mm,1.7μm),以流动相A水-甲醇-乙腈(1∶1∶1,V/V)和流动相B异丙醇-乙腈(1∶1,V/V)进行梯度洗脱,在电喷雾电离正模式下采集数据。依据高分辨质谱提供的保留时间、准分子离子、二级碎片等脂质分析数据,并结合标准品碎裂方式等信息,共鉴定出磷脂分子68种。其中,磷脂酰胆碱22种,占比7.98%;磷脂酰乙醇胺17种,占比56.60%;磷脂酰丝氨酸10种,占比1.70%;磷脂酰肌醇7种,占比1.33%;鞘磷脂12种,占比32.39%。本研究采用灵敏度高、精密度及稳定性良好的UPLC-Triple-TOF MS/MS方法实现了对牛乳中磷脂的全面定性与定量分析。此方法检测出的牛乳磷脂信息完整、磷脂种类多于其他检测技术,可为牛乳的磷脂研究提供数据基础。The phospholipids in pasteurized milk samples were analyzed and identified by ultra-high performance liquid chromatography-triple quadrupole-time of flight tandem mass spectrometry(UPLC-Triple-TOF MS/MS).The chromatographic separation was performed on an ACQUITY UPLC^®BEH C18(2.1 mm×100 mm,1.7μm)column using gradient elution with water/methanol/acetonitrile(1:1:1,V/V)as mobile phase A and isopropanol/acetonitrile(1:1,V/V)as mobile phase B.Data acquisition was done in the positive ion mode using electrospray ionization(ESI).According to the high-resolution mass spectral data of retention time,quasi-molecular ions and secondary fragments combined with information on the mass spectral fragmentation pathways of reference standards,a total of 68 phospholipid molecules were identified including 22 phosphatidylcholine(PC,7.98%),17 phosphatidylethanolamine(PE,56.60%),10 phosphatidylserine(PS,1.70%),7 phosphatidylinositol(PI,1.33%),and 12 sphingomyelin(SM,32.39%).This study achieved comprehensive qualitative and quantitative determination of phospholipids in milk by using the UPLC-Triple-TOF MS/MS method with high sensitivity,precision and stability.This method could detect more phospholipids in milk than other detection techniques and thus provide more basic data for research on milk phospholipids.

关 键 词：巴氏杀菌乳 磷脂 磷脂组学 超高效液相色谱-串联四极杆飞行时间质谱 

分 类 号：TS252.42[轻工技术与工程—农产品加工及贮藏工程]